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AIM: To observe the effect of Chinese medicine compound JiSuiKang on the microglia infiltration and neuron survival at the spinal cord injury area of rats. METHODS: Sixty SD rats were randomly divided into sham operation group (Sham), model group (Ctrl), JSK low (JSK-L), JSK medium (JSK-M) and JSK high (JSK-H) dose group, prednisone group. The model of rat spinal cord injury (SCI) was established by improved Allen's method. After the operation, each group was intervened in different ways, and then we assessed the double hind limb motor function score in rats (BBB score). The heart perfusion was done and the spinal cord tissues from the injury area were taken. ELISA tests were used to detect the expression of inflammatory factors. The spinal cord tissue was sliced into sections and then immunofluorescence stained of CD11b/c and beta 3-Tubulin to observe the infiltration of microglia and the growth of neurons under the inverted fluorescence microscope. RESULTS:The BBB score of Sham, JSK-L, JSK-M, JSK-H and prednisone group were higher than Ctrl(P0.05). For the growth state of the neuron, the JSK-L, JSK-M, JSK-H, prednisone group were better than Ctrl (P<0.01), and JSK-H was better than JSK-L and JSK-M (P<0.05). CONCLUSION: The Chinese medicine compound JSK inhibits the migration and invasion of microglia in SCI, promotes the recovery of damaged neurons and motor function in SCI rats.
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Objective:To observe the effects of Chinese herbal compound'Jisuikang'on the phagocytosis of microglia and the regeneration of injured neurons in co-culture system.Methods: Prepared drug serum of 'Jisuikang′ and isolated and identified the primary neuron and microglia.The neuron cells were induced apoptosis by glutamic acid and the microglia cells were predisposed by drug serum of'Jisuikang'.Then,the co-culture system of injured neurons and microglia cells was established.24 h and 96 h after co-culture,engulfment of neuron debris by microglia cells and regeneration of injured neurons were observed by immunofluorescence double labeling method.Results: 24 h after co-culture,middle and high dose of'Jisuikang' showed greater phagocytic percentage and phagocytic index than that of control.In comparison of LPS,high dose of'Jisuikang' showed no significant difference.96 h after co-culture,first grade of neuritis of middle and high dose of'Jisuikang' were more than that of control,and there were no significant difference in comparison of LPS.Neuritis' mean length per cell of middle and high dose of'Jisuikang' were larger than that of con-trol.Neuritis' mean length per cell of high dose of'Jisuikang' showed significant difference in comparison of LPS.Conclusion:Traditional Chinese medicine compound'Jisuikang'may enhance engulfment of neuron debris by microglia to improve local microenvi-ronment,which promote the repair and regeneration of injured neurons.
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This study was aimed to investigate the effect of traditional Chinese medicine (TCM) compound "Ji-Sui-Kang (JSK)" on the differentiation of neural stem cells (NSCs).Drug serum of JSK was prepared.The primary NSCs were isolated and cultured.NSCs were identified.Fetal bovine serum (FBS) was used to induce differentiation of NSCs.Different doses of drug serum of JSK was added to intervene the differentiation.There were the high-,medium-,and low-dose group.The blank control group was also set.The β Ⅲ-tubulin and GFAP were used to detect the differentiation of NSCs.And then,it was compared with the control group without drug serum.Positive cells ofβ m-tubulin and GFAP in different visual fields were counted.And the percentage of positive cells in the total number of cells was calculated.Fluorescence microscope was used to conduct quantitative fluorescence experiments.The results showed that the cultured neurospheres were expressed of Nestin protein.The percentage ofβⅢ-tubulin positive cells in the high-,medium-and low-dose group of drug serum of JSK was significantly higher than that in the control group (P＜0.05).The percentage of GFAP positive cells in the high-,medium-and low-dose groups of drug serum of JSK was significantly lower than that in the control group (P＜0.05).And the high-dose group of drug serum of JSK was statistically significant compared with the middle-and low-dose group of drug serum of JSK (P＜0.01).It was concluded that TCM compound JSK promoted thedifferentiation of NSCs intoβⅢ-tubulin positive cells.It provided a basis for treatment of SCI with TCM.
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Objective To clarify the effects of Jisuikang on expression levels of BDNF and NGF protein in serum of rats with spinal cord injury;To explore its probable mechanism to improve the axon regeneration microenvironment and resistance spinal cord injury.Methods Ninety SD rats were chosen to establish acute spinal cord injury model through modified Allen's method. 15 rats were chosen randomly as sham-operation group, and the other 75 rats were randomly divided into model group, prednisone group and high-, medium-, and low-doseJisuikang groups. All administration groups were given relevant medicine for gavage, while rats in sham-operation group and model group were given normal saline with the same volume for gavage. The activity and death condition of rats were recorded. Rats were put to death on the 3rd, 7th and 14th d of intervention for carotid artery blood collection. The expressions of BDNF and NGF in serum were detected by ELISA.Results After modeling, the rats in model group showed a typical paraplegia syndrome. 3 day after operation, BDNF showed no statistical significance between model group and prednisone group, while each treatment group showed significant difference when compared with prednisone group (P<0.05,P<0.01). 7 d and 14 d after operation, compared with the model group, BDNF of prednisone group and medium-dose Jisuikang group showed statistical significance, and there was no significant difference between prednisone group and medium-dose Jisuikang group (P<0.05,P<0.01). The expression of NGF in each treatment group at each time point with the prednisone group showed statistical significance. 3 d and 7 d after operation, among prednisone group, high- and medium-doseJisuikang groups, the expression of NGF was significantly higher than model group. 14 d after operation, compared with the model group, the expression of NGF in medium- and low-dose Jisuikang groups and prednisone group still maintained at a relatively high level, with statistical significance (P<0.05,P<0.01).Conclusion Jisuikang can effectively promote the expressions of BDNF and NGF in serum after spinal cord injury and maintained at a relatively high level, so as to improve the microenvironment of axonal regeneration and promote nerve regeneration.
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Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic fac-tor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away on-ly. The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg?d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg?d) Jisuikang respectively, which were given 20 ml/(kg?d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoper-ative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P0.05). The results of RQ-PCR showed that prednisone and Jisuikang promot-ed the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.